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Comparison to Methods; Serum Antibody ELISA and Fecal Nested-PCR to Diagnose Mycobacterium avium Subspecies Paratuberculosis Subspecies Infection in Cattle
2023
Kolivand, Ali | Haji Hajikolaei, Mohammad Rahim | Nouri, Mohammad | Khosravi, Mohammad | Gharibi, Dariush
BACKGROUND: Mycobacterium avium subspecies paratuberculosis is the cause of a common disease in dairy herds. Early diagnosis of paratuberculosis infection can improve Johne’s disease control programs.OBJECTIVES: This study aimed to compare the sensitivity, and specificity to methods; blood serum ELISA and stool Nested-PCR for the detection of Mycobacterium avium subspecies paratuberculosis infection in dairy cattle.METHODS: A commercial ELISA kit was used to perform the absorbed ELISA test, which was conducted after exposing serum samples to Mycobacterium phlei antigens to limit cross-reactions. Nested-PCR test was performed using nucleotide sequences related to specific MAP gene fragments, i.e. IS900.RESULTS: As a result of the ELISA antibodies kit, out of the total 2203 serum samples, 112 samples were positive (5.08 %) and 2091 samples were negative (94.92 %). The results of Nested-PCR tests of rectal feces showed that out of 59 cows with the positive results in serum ELISA, 47 (79.66 %) samples were positive and 12 (20.34 %) samples were negative. Moreover, out of 31 cattle with a negative result on the ELISA test, 15 (48.38%) and 16 cattle (51.62 %) had positive and negative results, respectively, on the nested PCR tests of the feces samples.CONCLUSIONS: Due to the low sensitivity of PCR compared to ELISA, the positive and negative predictive values, and the accuracy of ELISA test, as well as the high cost and time-consuming nature of PCR and the need for more and more complex facilities than ELISA, the authors concluded that ELISA is a more suitable method for screening and epidemiological studies than PCR.
Показать больше [+] Меньше [-]Prevalence Study, Pathological, Hematological, and Electrocardiographic Indices of Abomasal Ulcers at Sanandaj Abattoir
2021
Abaszadeh, Mohammad Sina | Fakour, Shahin | Akradi, Loghman
BACKGROUND: Abomasal ulcer is a disease of digestive system in ruminants. The clinical signs vary widely depending on the type of abomasal ulcer. The economic losses of this disease include decreased milk yield and mortality. OBJECTIVES: The present study aimed to determine the frequency, type of ulcer, pathological, hematological, and electrocardiographic indices in the slaughtered cattle in Sanandaj abattoir. METHODS: A total of 400 cattle were randomly selected with both sexes and 1-2, 2-3, 3-4, and ≥4 years of age. They were grouped during three seasons and the abomasal ulcer was evaluated. Blood sample was collected and electrocardiogram was recorded before they were slaughtered. Abomasal lesions were assessed macroscopically and histopathologically in the slaughtered cattle. RESULTS: Out of the 400 abomasom examined, 51 (12.75 %) were affected by abomasal ulcers, all of which were type-1 ulcer. Subtypes 1a, 1b, 1c, and 1d. 35 cattle (68.7 %) had the types of arrhythmia. The most frequent of them was sinus arrhythmia and sinus tachycardia. Neutrophilia and reduction in MCHC were significantly more than the other hematological indices (p < /em>˂0.05). There was a significant relationship between the prevalence of abomasal ulcers and arrhythmia, gender, age, and season (p < /em>˂0.05). Furthermore, there was a significant relationship between electrocardiographic and hematological indices in the affected cattle (p < /em>˂0.05). CONCLUSIONS: The results of this study revealed a high prevalence of abomasal ulcer in the studied area. They also indicated that the electrocardiographic and hematological changes could be helpful in the diagnosis of abomasal ulcer.
Показать больше [+] Меньше [-]Study on Cryptosporidium Infection in Cattle and Water Buffaloes of Farms in Mahabad Suburb, Iran
2020
Noorani Kolije, Omid | Yakhchali, Mohammad | Malekifard, Farnaz
BACKGROUND: The protozoancryptosporidiumisan important intestinal parasitic infection in domestic ruminants that has the potential for transmission between humans and livestock throughout the world and Iran. OBJECTIVES: The present study was undertaken to determine cryptosporidiuminfection in different age groupsof cattle and water buffaloesin farms of Mahabad suburb, Iran. METHODS: For this purpose, a total number of 248 fresh fecal samples were randomly collected from rectum of cattle and water buffaloesin farms of 4 villages from May2016 to May 2017. The fecal samples were subjected to floatation technique andcryptosporidiumoocysts were collected using Sucrose Gradient and Percole flotation technique and stained with modified Ziehl-Neelsen technique. RESULTS: Overall prevalence of cryptosporidiuminfection was 50% (124/248). The highest rate of infection was significant in female calves (30.65%) less than one year-old. The highest infection rate was significantly found in summer (20.16%). Cryptosporidium parvum and C. andersoni were identified in 40.32% (100/248) and 9.68% (24/248) of examined cattle, respectively. Mixed infection was 8.47%. CONCLUSIONS: The results of this study indicated that C.parvum was prevalent in cattle of the region, therefore, further molecular studies are recommended to determine the genotypes of the parasiteas a potential zoonotic agent.
Показать больше [+] Меньше [-]Antibacterial Effect of Olive Leaf (Olea europaea. L) Alcoholic Extract on Staphylococcus aureus Isolated from Milk Sample of Cows with Subclinical Mastitis
2022
Shakiba, Hossein | Mohammadi, Hamid Reza | Jebelli Javan, Ashkan | Narenji Sani, Reza
BACKGROUND: Subclinical mastitis plays an important role in the economic losses of dairy cattle farms. Staphylococcus aureus is one of the most important causes of this disease. Treatment of this disease with synthetic antibiotics has complications like antibiotic resistance. Using herbal antibiotics can be an excellent way to reduce these side effects.OBJECTIVES: This study aimed to determine the minimum inhibitory concentration of alcoholic extract of olive leaf on Staphylococcus aureus isolated from milk of cows with subclinical mastitis to achieve herbal treatment.METHODS: This study was conducted on 175 Holstein female cattle. The milk samples of 60 cows were obtained with the sterilized method, and Subclinical mastitis-positive cases were determined using the California mastitis test. Staphylococcus aureus bacteria were isolated from positive samples by culture method, and the minimum inhibitory concentration of olive (Olea europaea L.) leaves alcoholic extract on isolated bacteria was determined by microdilution method.RESULTS: From 175 cows under study, 60 cows had a positive California mastitis test, and Staphylococcus aureus separated from milk samples of 14 cows. The minimum inhibitory concentration of olive (Olea europaea L.) leaves extract on this bacterium was 12000 ppm.CONCLUSIONS: Alcoholic extract of olive (Olea europaea L.) leaves has an antimicrobial effect on Staphylococcus aureus as a cause of mastitis. The minimum concentration required for this effect was 12000 ppm. Further studies on the impact of this plant on other bacterial causes of subclinical mammary inflammation in cows and investigation of the effective substances in the extract are needed.
Показать больше [+] Меньше [-]Slaughterhouse Survey of Cobalt Status in Serum and Liver of Cattle in Different Seasons
2019
Bahrami, Ahmad | Asri-Rezaei, Siamak | Akbari, Hamid | Dalir-Naghadeh, Bahram
BACKGROUND: Cobalt is an essential nutrient for ruminants which is required for the synthesis of vitamin B12 by the ruminal microflora. Cobalt deficiency can lead to a deficiency of vitamin B12, which plays a major role in many metabolic reactions in the body. OBJECTIVES: This study was conducted to evaluate cobalt status in the serum and liver of cattle. METHODS: In different seasons, the blood serum and liver concentration of cobalt in slaughtered cattle in Urmia city were measured by atomic absorption in 151 and 196 cattle, respectively. RESULTS: The results of cobalt measurement indicated that in 12.6% of serum and 21.4% of liver samples the concentration of cobalt was less than normal range with the Mean ± SD value of 0.64 ± 0.25 µg/dl and 0.15 ± 0.04 µg/gDM, respectively. The seasonal variations of cobalt values were assessed in serum and liver samples and following results were obtained: spring (2.18 ± 1.64 µg/dl, 0.23 ± 0.13 µg/gDM), summer (3.42 ± 1.04 µg/dl, 0.42 ± 0.27 µg/gDM), autumn (3.45 ± 0.37 µg/dl, 0.47 ± 0.38 µg/gDM) and winter (2.75 ± 0.59 µg/dl, 0.35 ± 0.09 µg/gDM). CONCLUSIONS: According to the results of this study, it seems that there is a subclinical deficiency of cobalt in cattle of this region and it is recommended that preventive measures be considered to prevent the complications and disorders caused by its clinical deficiency.
Показать больше [+] Меньше [-]Seroprevalence of Peste Des Petits Ruminants (PPR) Virus Infection in Sheep and Cattle in Ahvaz
2018
Rasooli, Aria | Nouri, Mohammad | Seyfi Abadshapouri, Masoud-Reza | Khalafi, Elham | Daghari, Maryam
BACKGROUND: Peste des petits ruminants (PPR) is an acute and highly contagious viral disease of small ruminants that is characterized by high fever, ocular and nasal discharge, pneumonia, necrosis, ulceration of the mucous membranes and inflammation of the gastro-intestinal tract leading to severe diarrhea. OBJECTIVES: The aim of this study was to investigate the seroprevalence of peste des petits ruminants (PPR) virus infection in sheep and cattle in Ahvaz. METHODS: Blood samples were taken from 100 cattle and 100 sheep that were kept together from different parts of Ahvaz. Blood samples were also taken from 16 vaccinated sheep against PPR for positive control. The sera were separated by centrifuge at 3000 ×g for 10 minutes and 3 mL of serum was harvested and stored at -20 °C until determination of antibody against PPR using VN method. RESULTS: The peste des petits ruminants (PPR) antibody seroprevalence was 23% in cattle and 58% in sheep and all the sheep samples collected for control were positive for PPR antibody. CONCLUSIONS: The present study indicates serological evidence for the natural transmission of PPRV from sheep to cattle under natural conditions and provides baseline information on prevalence of PPRV antibodies in cattle and sheep population in Ahvaz.
Показать больше [+] Меньше [-]Molecular detection of Theileria species and its vectors in cattles in Yazd area by semi-nested PCR method
2015
Khodabandeh, Soghra | Razmi, Gholamreza
BACKGROUND: Theileriosis is a blood protozoan disease with high mortality in cattle in tropical and subtropical regions of the world. Several studies were conducted to identify Theileria species in infected cattles and vector ticks by microscopic examination. However, microscopic technique has lower sensitivity compared to molecular method. OBJECTIVES: This study was carried out to identify Theileria species and its carriers in cattles of Yazd city, sing semi nested PCR. METHODS: Between June to September of 2012, 100 EDTA blood samples and 249 ticks were collected from Holstein breed with no history of vaccination against the ileriosis in Yazd area. The collected samples were transported to the laboratory, then prepared the blood smears and stained with Giemsa method. Also, the collected ticks were separated into 50 tick pools, according to their species. Then their salivary glands were removed using stereomicroscope in 0.85% saline. DNA of blood and salivary glands was extracted using a commercial kit and analyzed by Semi-nested PCR. PCR-RFLP was also used to differentiate Theileria lestoquardi from Theileria annulata in positive samples of ticks. RESULTS: Ring forms of Theileria spp. were found in 4 (4%) of blood smears. All ticks were Hyalomma a.anatolicum. Results of PCR were indicated that 11 (11%) of blood samples and three pools of tick’s salivary glands were infected with Theileria. Theileria annulata were only detected in all positive samples by Semi-nested PCR. The results of PCR-RFLP using MspI enzyme showed that the tick’s salivary glands were negative to Theileria lestoquardi infection. CONCLUSIONS: Based on the results, it is concluded that Theileria annulata and Hyalomma a.anatolicum are important agent and vector tick of tropical theileriosis in dairy cattles of Yazd area.
Показать больше [+] Меньше [-]Hard ticks (Acari: Ixodidae) Infestation in Ruminants of Some Areas in Ilam Province, Iran
2019
Ghashghaei, Omid | Yakhchali, Mohammad | Nourollahi-Fard, Saeed Reza
BACKGROUND: Ticks are important ectoparasites in ruminants which cause economic losses in animal husbandry of Iran and worldwide. OBJECTIVES: This study was aimed to determine geographic distribution, frequency and species diversity of hard ticks in domestic ruminants in Ilam province, Iran. METHODS: A total of 445 domestic ruminants (139 cattle, 162 sheep, 144 goats) from 120 flocks of 30 villages in north and south parts of Ilam province were randomly selected and examined in summer 2015. The ixodid ticks were collected from body surface of examined animals and identified. RESULTS: Of all examined ruminants, 44.6% cattle, 51.23% sheep, and 52.08% goats were infested with a total number of 1209 unfed ixodid ticks. The highest prevalence of hard ticks was found in 2-3 year-old female cattle (15.55%), 1-2 year-old female sheep (19.75%) and less than 1 year-old female goats (17.36%) in the region. The highest prevalence was found in 2-3 year-old female cattle (37.1%), 1-2 year-old ewes (38.55%) and less than 1 year-old kids (33.33%). There was significant difference between prevalence and different age groups of infested sheep and cattle. Of 1209 collected ixodid ticks, tick indices (tick number per animal) were 6.1, 5.9 and 4.5 in cattle, sheep, and goats, respectively. The highest infestation in cattle (17 flocks, 14.1%), sheep (13 flocks, 10.8%) and goats (14 flocks, 11.6%) was respectively found in north, south and southern parts of the region. The highest tick aggregation was found for ears in cattle (31.13%), sheep (34.41%) and goats (28.9%). Of all examined ticks (1209), two genera including Hyalomma (37.62%) and Rhipicephalus (62.38%) with seven, seven, and six species in cattle, sheep, and goats were respectively identified. The predominant infesting ticks were R. sanguineus (22.16% in cattle) from north (11.31%) and H. anatolicum anatulicum (21.8% in sheep and 24.77% in goats) from south (15.49% and 13.42%) part of the province. CONCLUSIONS: The results revealed that species diversity and frequency of ixodid ticks were prevalent in domestic ruminants of different parts of Ilam province.
Показать больше [+] Меньше [-]Morphological and molecular characterization of cattle hydatid cysts in Khorramabad, Iran
2016
Kasaeiyan, Fatemeh | Nayebzadeh, Hassan | Jalousian, Fatemeh | Shokrani, Hamid Reza
BACKGROUND: Hydatidosis is one of the zoonotic diseases which affect animals and human beings at the larval stage of Echinococcusgranulosus, thereby playing a role in exacerbating the economic and health problems in Iran. OBJECTIVES: This study was conducted to investigate the strains of Echinococcusgranulosus isolated from cattle hydatid cysts, in Khorramabad, in the Lorestan Province, west of Iran. METHODS: Twenty six isolates of hydatid cyst of cattle from lung (24 samples) and liver (2 sample) organs were collected from Golshan Slaughterhouse, in Khorramabad. All of the samples were transferred to the laboratory for morphometric characterization and molecular study. In morphological characterization, blade length of large (LBL) and small (SBL) hook and the ratio of blade length to total length in large (LBL/LTL) and small (SBL/STL) hooks and total length of large (LTL) and small (STL) hooks were measured. In molecular study, a partial sequence of cytochrome oxidase 1 (COX1) with 440 bp in length was amplified applying primers J3 and J4.5. Genomic DNA sequencing was performed by Sanger’s method. RESULTS: The morphological results showed that there is no significant difference between isolated from cattle hydatid cyst and sensu stricto strain (P<0.05). The results of molecular studies support the findings of morphological characterization. All sequences showed 100% identity with sensu stricto strain. CONCLUSIONS: The results from this study showed that sensu stricto strain (G1) is a causative agent of cattle hydatid cyst in Khorramabad. On the other hand, the cattle play a role in enabling the parasite to complete its cycle. Hence in order to execute a control program for minimizing the effects of this disease, the cattle shall be considered as a source of infection for dogs. The results of this study could be helpful in designing such control program in the region.
Показать больше [+] Меньше [-]Influence of nocodazole on the development of donor blastomeres from 16-cell stage bovine embryos in nuclear transfer.
1995
Tanaka H. | Takahashi Y. | Hishinuma M. | Kanagawa H. | Kariya T.
The aim of the present study was to establish a reliable procedure with nocodazole treatment for the synchronous cleavage of blastomeres of bovine embryos used as nuclear donors for nuclear transfer. Sixteen-cell stage embryos derived from in vitro-maturation, fertilization and culture were used. In three initial experiments, embryos were incubated in mTCM-199 + FCS with various concentrations (0-20 mu-M) of nocodazole under 5% CO2 in air. The concentrations required to arrest the blastomeres in the mitotic phase were examined. The effects of 10 mu-M nocodazole were also examined by observation of the division rate of blastomeres after the removal of nocodazole. Ninety percent (90%) of the blastomeres were arrested in the mitotic phase when embryos were exposed to 10 and 20 mu-M nocodazole. Exposure to 10 mu-M nocodazole had the highest blastomere-cleavage rate (47%). When the exposure period to 10 mu-M nocodazole was prolonged to 36 hr, the division rate of the blastomeres decreased. Furthermore, the effects of 2 culture conditions (mTCM-199 under 5% CO2 in air vs modified synthetic oviduct fluid medium under 5% CO2, 5% O2 and 90% N2) were compared on the division rate of blastomeres of embryos exposed to 10 mu-M nocodazole for 12 hr. When the embryos were exposed to nocodazole in mSOF, the division rate of blastomeres was improved to about 60%. The blastomeres produced by this treatment condition were used as nuclear donors and the developmental potential of the reconstituted embryos was investigated. The developmental rate to the blastocyst stage was 30.1% (58/193). Five embryos were transferred to 5 recipient cows and 2 of the 5 recipients (40%) became pregnant. Subsequently, one normal calf was born.
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